Polyamine compounds for treating chemokine receptor mediated diseases

ABSTRACT

This invention relates to a method for treating inflammatory diseases or immune diseases, developmental or degenerative diseases, or tissue injuries. The method includes administering to a subject in need thereof an effective amount of one or more compounds of the following formula. Each variable in this formula is defined in the specification.

CROSS REFERENCE TO RELATED APPLICATION

Pursuant to 35 USC § 119(e), this application claims priority to U.S.Provisional Application Ser. No. 60/459,768, filed Apr. 2, 2003, andU.S. Provisional Application Ser. No. 60/539,763, filed Jan. 28, 2004,the contents of which are incorporated herein by reference.

BACKGROUND

Chemokines are a family of cytokines that regulate the adhesion andtransendothelial migration of leukocytes during an immune orinflammatory reaction (Mackay C. R., Nat. Immunol., (2001) 2:95; Olsonet al., Am. J. Physiol. Regul. Integr. Comp. Physiol., (2002) 283:R7).Chemokines also regulate T cells and B cells trafficking and homing, andcontribute to the development of lymphopoietic and hematopoietic systems(Ajuebor et al., Biochem. Pharmacol., (2002) 63:1191). Approximately 50chemokines have been identified in humans. They can be classified into 4subfamilies, i.e., CXC, CX3C, CC, and C chemokines, based on thepositions of the conserved cysteine residues at the N-terminal (Onufferet al., Trends Pharmacol Sci., (2002) 23:459). The biological functionsof chemokines are mediated by their binding and activation of Gprotein-coupled receptors (GPCRs) on the cell surface. Take CXCR4receptor for example, it can be activated by Stromal-derived factor-1 orSDF-1, a member of CXC chemokines.

SDF-1 was originally cloned from bone marrow stromal cell lines andfound to act as a growth factor for progenitor B cells (Nishikawa etal., Eur. J. Immunol., (1988) 18:1767). SDF-1 also induces bone marrowcolonization of hematopoietic precursor cells during embryogenesis(Bleul et al., J. Exp. Med., (1996) 184:1101). The physiologicalfunction of SDF-1 is mediated by CXCR4 receptor. Mice lacking SDF-1 orCXCR4 receptor show lethal abnormality in bone marrow myelopoiesis, Bcell lymphopoiesis, and cerebellar development (Nagasawa et al., Nature,(1996) 382:635; Ma et al., Proc. Natl. Acad. Sci., (1998) 95:9448; Zouet al., Nature (1998) 393:595; Lu et al., Proc. Natl. Acad. Sci. (2002)99:7090). CXCR4 receptor is expressed broadly in a variety of tissues,particularly in immune and central nervous systems, and has beendescribed as the major co-receptor for HIV-1/2 on T lymphocytes.Although initial interest in CXCR4 antagonism focused on its potentialapplication to AIDS treatment (Bleul et al., Nature (1996) 382:829), itis now becoming clear that CXCR4 receptor and SDF-1 are also involved inother pathological conditions such as rheumatoid arthritis, asthma, andtumor metastases (Buckley et al., J. Immunol., (2000) 165:3423). CXCR4receptor and SDF-1 are also found widely expressed in many tissuesduring embryonic development. Further, the CXCR4/SDF-1 pathway has beenshown to be critically involved in the regeneration of several tissueinjury models. Specifically, it has been found that the SDF-1 level iselevated at an injured site and CXCR4-positive cells activelyparticipate in the tissue regenerating process.

SUMMARY

This invention is based on the discovery that certain polyaminecompounds are effective in treating inflammatory and immune diseasesthrough their binding to chemokine receptors (e.g., CXCR3 or CXCR4receptors).

In one aspect, this invention features polyamine compounds of theformula:

In the above formula, X is —CH₂—, —C₂H₄—, —C₃H₆—, —CH₂—CH═CH—,—CH═CH—CH₂—, —C(O)—, —SO₂—, or deleted; Y is aryl, heteroaryl, C₃-C₈cycloalkyl, C₅-C₈ cycloalkenyl, C₃-C₈ heterocycloalkyl, or C₅-C₈heterocycloalkenyl; each of Z₁ and Z₂, independently, is —CH₂—, —C₂H₄—,—C₃H₆—, —CH═CH—, —CH═N—, —CH═N—NR—, —S—, —O—, —NR—, —C(O)—, or —SO₂—; R₁is H, C₁-C₁₀ alkyl, C₂-C₁₀ alkenyl, C₂-C₁₀ alkynyl, C₃-C₈ cycloalkyl,C₅-C₈ cycloalkenyl, C₃-C₈ heterocycloalkyl; C₅-C₈ heterocycloalkenyl,aryl, or heteroaryl; R₂ is -A₁-B₁-D₁-E₁; R₃ is -A₂-B₂-D₂-E₂, deleted,or, together with R₄, is C₄-C₂₀ cycloalkyl, C₄-C₂₀ cycloalkenyl, C₄-C₂₀heterocycloalkyl, or C₄-C₂₀ heterocycloalkenyl; provided that if R₃ isdeleted, -Z₂-N— is —CH═N—; and R₄ is -A₃-B₃-D₃-E₃ or, together with R₃,is C₄-C₂₀ cycloalkyl, C₄-C₂₀ cycloalkenyl, C₄-C₂₀ heterocycloalkyl, orC₄-C₂₀ heterocycloalkenyl. Each of A₁, A₂, and A₃, independently, is—CH₂—, —C₂H₄—, —C₃H₆—, —C₄H₈—, —C₅H₁₀—, —CH₂C(O)—, —C(O)CH₂—, —CH₂SO₂—,—SO₂CH₂—, —CH₂—CH═CH—, —CH═CH—CH₂—, —CH(CH₂OR)—, —CH(CH₂CH₂OR)—,—CH(COOR)—, —CH(CH₂COOR)—, —CH(C(O)NR₂)—, or deleted. Each of B₁, B₂,and B₃, independently, is —NR—, —CH₂—, or deleted. Each of D₁, D₂, andD₃, independently, is —CH₂—, —C₂H₄—, —C₃H₆—, —CH₂—CH═CH—, —CH═CH—CH₂—,—C(O)—, —SO₂—, —C(O)—NR—, —C(S)—NR—, —NR—C(O)—, —NR—C(S)—, —CH(OR)—,—CH(CH₂OR)—, —CH(CH₂CH₂OR)—, —CH(COOR)—, 1,1-cyclopropylene, or deleted.Each of E₁, E₂, and E₃, independently, is H, C₁-C₁₀alkyl, C₂-C₁₀alkenyl, C₂-C₁₀ alkynyl, C₃-C₈ cycloalkyl, C₅-C₈ cycloalkenyl, C₃-C₈heterocycloalkyl, C₅-C₈ heterocycloalkenyl, aryl, or heteroaryl. Each R,independently, is H or C₁-C₁₀ alkyl.

Referring to the above formula, a subset of the just-described compoundsare those in which X is —CH₂— or —CH(CH₃)—, Y is phenyl or4,4′-biphenyl, Z₁ is —CH₂— or —SO₂—, Z₂ is —CH₂— or —SO₂—, or R₃ isdeleted.

The term “alkyl” refers to a saturated, linear or branched, non-aromatichydrocarbon moiety, such as CH₃, —CH₂—, or branched C₃H₇. Referring tothe above formula, —C₂H₄— and —C₃H₆— can be either linear or branched.The term “alkenyl” refers to a linear or branched, non-aromatichydrocarbon moiety having at least one double bond, such as —CH═CH₂ or—CH═CH—. The term “alkynyl” refers to a linear or branched, non-aromatichydrocarbon moiety having at least one triple bond, such as —C≡CH or—C≡C—. The term “cycloalkyl” refers to a saturated non-aromatic cyclichydrocarbon moiety. The term “cycloalkenyl” refers to a non-aromaticcyclic hydrocarbon moiety having at least one double bond in the ring.The term “heterocycloalkyl” refers to a saturated non-aromatic cyclicmoiety having at least one ring heteroatom (e.g., O, N, and S). The term“heterocycloalkenyl” refers to a non-aromatic cyclic moiety having atleast one ring heteroatom and at least one double bond in the ring. Theterm “aryl” refers to a hydrocarbon moiety having at least one aromaticring. Examples of an aryl moiety include phenyl, phenylene, biphenyl,naphthyl, naphthylene, pyrenyl, anthryl, and phenanthryl. The term“heteroaryl” refers to a moiety having at least one aromatic ring whichcontains at least one heteroatom. Examples of a heteroaryl moietyinclude furyl, furylene, fluorenyl, pyrrolyl, thienyl, oxazolyl,imidazolyl, thiazolyl, pyridyl, pyrimidinyl, quinazolinyl, and indolyl.

Alkyl, alkenyl, alkynyl, cycloalkyl, cycloalkenyl, heterocycloalkyl,heterocycloalkenyl, aryl, and heteroaryl mentioned herein include bothsubstituted and unsubstituted moieties. Examples of substituents forcycloalkyl, cycloalkenyl, heterocycloalkyl, heterocycloalkenyl, aryl,and heteroaryl include C₁-C₁₀ alkyl, C₂-C₁₀ alkenyl, C₂-C₁₀ alkynyl,C₃-C₈ cycloalkyl, C₅-C₈ cycloalkenyl, C₁-C₁₀ alkoxy, aryl, aryloxy,heteroaryl, heteroaryloxy, amino, C₁-C₁₀ alkylamino, C₁-C₂₀dialkylamino, arylamino, diarylamino, heteroarylamino,diheteroarylamino, C₁-C₁₀ alkylsulfonyl, arylsulfonyl,heteroarylsulfonyl, C₁-C₁₀ alkylsulfonamide, arylsulfonamide,heteroarylsulfonamide, hydroxyl, halogen, mercapto, C₁-C₁₀alkylmercapto, arylmercapto, cyano, nitro, acyl, acyloxy, carboxyl,amido, carbamoyl, and carboxylic ester. Examples of substituents foralkyl, alkenyl, and alkynyl include all of the above substitutentsexcept C₁-C₁₀ alkyl, C₂-C₁₀ alkenyl, and C₂-C₁₀ alkynyl. Cycloalkyl,heterocycloalkyl, aryl, and heteroaryl also include fused groups.

In another aspect, this invention features polyamine compounds of thesame formula shown above. Referring to this formula, the same groups asthose described above are assigned to each variable except that X is—CH₂—, —C₂H₄—, —C₃H₆—, —CH₂—CH═CH—, —CH═CH—CH₂—, —SO₂—, or deleted; Y isaryl, heteroaryl, C₃-C₈ cycloalkyl, C₅-C₈ cycloalkenyl, C₃-C₈heterocycloalkyl, C₅-C₈ heterocycloalkenyl, or deleted; each of D₁, D₂,and D₃, independently, is —CH₂—, —C₂H₄—, —C₃H₆—, —CH₂—CH═CH—,—CH═CH—CH₂—, —SO₂—, —C(O)—NR—, —C(S)—NR—, —NR—C(O)—, —NR—C(S)—,—CH(OR)—, —CH(CH₂OR)—, —CH(CH₂CH₂OR)—, —CH(COOR)—, 1,1-cyclopropylene,or deleted; and E₁ is H, C₁-C₁₀ alkyl, C₂-C₁ alkenyl, C₂-C₁ alkynyl,C₃-C₈ cycloalkyl, C₅-C₈ cycloalkenyl, C₃-C₈ heterocycloalkyl, C₅-C₈heterocycloalkenyl, aryl, 5-membered heteroaryl, fused heteroaryl,substituted 6-membered heteroaryl, unsubstituted pryanyl, unsubstitutedpyrazinyl, unsubstituted pyrimidinyl, or unsubstituted pyridazinyl.

Referring to the above formula, a subset of the just-described compoundsare those in which X is —CH₂— or —CH(CH₃)—, Y is deleted, Z₁ is —CH₂—,or Z₂ is —CH₂—.

In still another aspect, this invention features a method for treatingan inflammatory or immune disease, a developmental or degenerativedisease, or a tissue injury. The method includes administering to asubject in need thereof an effective amount of one or more compounds ofthe same formula shown above. Referring to this formula, X is —CH₂—,—C₂H₄—, —C₃H₆—, —CH₂—CH═CH—, —CH═CH—CH₂—, —C(O)—, —SO₂—, or deleted; Yis aryl, heteroaryl, C₃-C₈ cycloalkyl, C₅-C₈ cycloalkenyl, C₃-C₈heterocycloalkyl, C₅-C₈ heterocycloalkenyl, or deleted; each of Z₁ andZ₂, independently, is —CH₂—, —C₂H₄—, —C₃H₆—, —CH═CH—, —CH═N—, —CH═N—NR—,—S—, —O—, —NR—, —C(O)—, or —SO₂—; R₁ is H, C₁-C₁₀ alkyl, C₂-C₁₀ alkenyl,C₂—C₁₀ alkynyl, C₃-C₈ cycloalkyl, C₅-C₈ cycloalkenyl, C₃-C₈heterocycloalkyl; C₅-C₈ heterocycloalkenyl, aryl, or heteroaryl; R₂ is-A₁-B₁-D₁-E₁; R₃ is -A₂-B₂-D₂-E₂, deleted, or, together with R₄, is C₂₀cycloalkyl, C₄-C₂₀ cycloalkenyl, C₄-C₂₀ heterocycloalkyl, or C₄-C₂₀heterocycloalkenyl; provided that if R₃ is deleted, -Z₂-N— is —CH═N—;and R₄ is -A₃-B₃-D₃-E₃ or, together with R₃, is C₄-C₂₀ cycloalkyl,C₄-C₂₀ cycloalkenyl, C₄-C₂₀ heterocycloalkyl, or C₄-C₂₀heterocycloalkenyl. Each of A₁, A₂, and A₃, independently, is —CH₂—,—C₂H₄—, —C₃H₆—, —C₄H₈—, —C₅H₁₀—, —CH₂C(O)—, —C(O)CH₂—, —CH₂SO₂—,—SO₂CH₂—, —CH₂—CH═CH—, —CH═CH—CH₂—, —CH(CH₂OR)—, —CH(CH₂CH₂OR)—,—CH(COOR)—, —CH(CH₂COOR)—, —CH(C(O)NR₂)—, or deleted. Each of B₁, B₂,and B₃, independently, is —NR—, —CH₂—, or deleted. Each of D₁, D₂, andD₃, independently, is —CH₂—, —C₂H₄—, —C₃H₆—, —CH₂—CH═CH—, —CH═CH—CH₂—,—C(O)—, —SO₂—, —C(O)—NR—, —C(S)—NR—, —NR—C(O)—, —NR—C(S)—, —CH(OR)—,—CH(CH₂OR)—, —CH(CH₂CH₂OR)—, —CH(COOR)—, 1,1-cyclopropylene, or deleted.Each of E₁, E₂, and E₃, independently, is H, C₁-C₁₀ alkyl, C₂-C₁₀alkenyl, C₂-C₁₀ alkynyl, C₃-C₈ cycloalkyl, C₅-C₈ cycloalkenyl, C₃-C₈heterocycloalkyl, C₅-C₈ heterocycloalkenyl, aryl, or heteroaryl. Each R,independently, is H or C₁-C₁₀ alkyl.

For example, one can administer to a subject having an above-describeddisease a compound of the above formula, in which X is —CH₂— or—CH(CH₃)—, Y is phenyl, 4,4′-biphenyl, or deleted, Z₁ is —CH₂— or —SO₂—,Z₂ is —CH₂— or —SO₂—, or R₃ is deleted.

“Treatment” refers to administering one or more polyamine compounds to asubject, who has an above-described disease, a symptom of such adisease, or a predisposition toward such a disease, with the purpose toconfer a therapeutic effect, e.g., to cure, relieve, alter, affect,ameliorate, or prevent the above-described disease, the symptom of it,or the predisposition toward it.

An inflammatory disease is characterized by a local or systemic, acuteor chronic inflammation. Examples include inflammatory dermatoses (e.g.,dermatitis, eczema, atopic dermatitis, allergic contact dermatitis,urticaria, necrotizing vasculitis, cutaneous vasculitis,hypersensitivity vasculitis, eosinophilic myositis, polymyositis,dermatomyositis, and eosinophilic fasciitis), inflammatory boweldiseases (e.g., Crohn's disease and ulcerative colitis),hypersensitivity lung diseases (e.g., hypersensitivity pneumonitis,eosinophilic pneumonia, delayed-type hypersensitivity, interstitial lungdisease or ILD, idiopathic pulmonary fibrosis, and ILD associated withrheumatoid arthritis), asthma, and allergic rhinitis.

An immune disease is characterized by a hyper- or hypo-reaction of theimmune system. Examples include autoimmune diseases (e.g., rheumatoidarthritis, psoriatic arthritis, systemic lupus erythematosus, myastheniagravis, juvenile onset diabetes, glomerulonephritis, autoimmunethroiditis, ankylosing spondylitis, systemic sclerosis, and multiplesclerosis), acute and chronic inflammatory diseases (e.g., systemicanaphylaxia or hypersensitivity responses, drug allergies, insect stingallergies, graft rejection, including allograft rejection, andgraft-versus-host disease), Sjogren's syndrome, human immunodeficiencyvirus infection, cancer (e.g., brain, breast, prostate, colon, kidney,ovary, thyroid, lung, and haematopoietic cancer), and tumor metastasis.

Developmental diseases are growth or differentiation related disordersthat lead to loss-of-function or gain-of-function. Degenerative diseasesgenerally refer to change of a tissue to a lower or less functionalform. Examples of a developmental or degenerative disease include spinalmuscular atrophy, Duchenne muscular dystrophy, Parkinson's disease, andAlzheimer's disease. Tissue injuries can be caused by oxidative stress(e.g., ischemia-reperfusion in stroke or myocardial infarction),complement activation, graft rejection, chemicals (e.g., alcohol-inducedliver damage or mucosal tissue injuries in cancer therapy), viralinfection (e.g., glomerular injuries associated with hepatitis Cinfection), and mechanical forces (e.g., sports injury). Examples oftissue injuries include brain injury, heart injury, liver damage,skeletal muscle injury, kidney damage, pancreatic injury, lung injury,skin injury, and gastrointestinal tract injury.

A subject in need of treatment of an above-described disease can also beconcurrently administered with a polyamine compound described above andone or more other therapeutic agents. Examples of such a therapeuticagent include a steroidal or a non-steroidal anti-inflammatory drug, aCOX2 inhibitor, a leukotriene receptor inhibitor, a prostaglandinmodulator, a TNF modulator, and an immunosuppressive agent (e.g.,cyclosporine A). The term “concurrently administered” refers toadministering a polyamine compound and one or more other therapeuticagents at the same time or at different times during the period oftreatment.

In a further aspect, this invention features a method for enhancingmigration of bone marrow-derived cells to blood. The method includesadministering to a subject in need thereof an effective amount of one ormore compounds of the same formula shown above. Referring to thisformula, the same groups as those described above are assigned to eachvariable. The term “bone marrow-derived cells” refers to cellsoriginating from bone marrow. Examples of bone marrow-derived cellsinclude, but are not limited to, CD34+ cells and CD133+ cells.

Also within the scope of this invention is a pharmaceutical compositionthat contains an effective amount of at least one of the above-mentionedpolyamine compounds and a pharmaceutically acceptable carrier.

The polyamine compounds described above include the compoundsthemselves, as well as their salts, prodrugs, and solvates, ifapplicable. A salt, for example, can be formed between an anion and apositively charged group (e.g., amino) on a polyamine compound. Suitableanions include chloride, bromide, iodide, sulfate, nitrate, phosphate,citrate, methanesulfonate, trifluoroacetate, and acetate. Likewise, asalt can also be formed between a cation and a negatively charged group(e.g., carboxylate) on a polyamine compound. Suitable cations includesodium ion, potassium ion, magnesium ion, calcium ion, and an ammoniumcation such as tetramethylammonium ion. The polyamine compounds alsoinclude those salts containing quaternary nitrogen atoms. Examples ofprodrugs include esters and other pharmaceutically acceptablederivatives, which, upon administration to a subject, are capable ofproviding active polyamine compounds. A solvate refers to a complexformed between an active polyamine compound and a pharmaceuticallyacceptable solvent. Examples of pharmaceutically acceptable solventsinclude water, ethanol, isopropanol, ethyl acetate, acetic acid, andethanolamine.

Also within the scope of this invention is a composition containing oneor more of the polyamine compounds described above for use in treatingan above-described disease, and the use of such a composition for themanufacture of a medicament for the just-mentioned treatment.

The details of one or more embodiments of the invention are set forth inthe description below. Other features, objects, and advantages of theinvention will be apparent from the description and from the claims.

DETAILED DESCRIPTION

Shown below are exemplary compounds, compounds 1-126, of this invention:

The polyamine compounds described in the summary section above can beprepared by methods well known in the art, including the syntheticroutes disclosed herein.

For example, compound 1-11 can be respectively prepared by reactingtris(2-aminoethyl)amine with three equivalent amounts of a correspondingaldehyde, and followed by a reduction reaction using sodium borohydride.

In another example, one can react 1,4-dibromoxylene with two equivalentamounts of bis(t-butoxycarbonylaminoethyl)amine. The reaction mixture issubsequently treated with hydrochloric acid to give an intermediate,1,4-di[bis(2-aminoethyl)aminomethyl]benzene. This intermediate can reactwith four equivalent amounts of corresponding aldehyde compounds andsodium borohydride to give compounds 12-14. Alternatively, thisintermediate can react with two equivalent amounts ofpyridine-2-carbaldehyde, and then with sodium borohydride to givecompound 15. One can also react 1,4-dibromoxylene with one equivalentamount of bis(t-butoxycarbonylaminoethyl)amine and one equivalent amountof other amine compounds. Compounds 56-57, 65, 66, 68, 80, 86, 91,93-95, and 106-109 can be prepared through this synthetic route,followed by sequential treatments with hydrochloric acid, two equivalentamounts of a corresponding aldehyde, and sodium borohydride. In asimilar manner, compounds 49 can be prepared by reacting1,4-dibromoxylene with one equivalent amount of tri-t-butoxycarbonylprotected cyclam and one equivalent amount ofbis(2-pyridiyliminoethyl)amine, followed by a reduction reaction usingsodium borohydride.

In another example, compounds 16-48, 58-64, 69-79, 81-85, 87-90, 92,96-105, 115, 121, 122, 125, and 126 can be synthesized using thefollowing synthetic route. One can react 4-cyanobenzylbromide with oneequivalent amount of bis(t-butoxycarbonylaminoethyl)amine and thenhydrochloric acid to give4-[bis(2-aminoethyl)amino-methyl]-benzonitrile. This benzonitrile canthen be treated sequentially with two equivalent amounts of an aldehyde,methyl]-benzaldehyde. The just-mentioned compounds can then be preparedby treating this benzaldehyde sequentially with one equivalent amount ofa corresponding amine, sodium borohydride, and hydrochloric acid.Compounds 110, 111, 116, 117, 120, 123, and 124 can be prepared in asimilar manner except that the benzonitrile is treated with oneequivalent amount of an aldehyde and one equivalent amount of a ketone.

In another example, one can react 4-bromomethylbenzenesulfonyl chloridewith one equivalent amount of an amine and then one equivalent amount ofbis(t-butoxycarbonylaminoethyl)amine. An intermediate is then obtainedafter treating the above reaction mixture with hydrochloric acid.Compounds 51-55 can then be respectively prepared by treating thisintermediate with a corresponding aldehyde and followed with sodiumborohydride. Compounds 50, 67, 112, and 113 can be prepared using thesame procedure as that of compounds 51-55 except that4-bromomethylbenzenesulfonyl chloride is treated with two equivalentamounts of bis(t-butoxycarbonylaminoethyl)amine.

In another example, one can react 4-cyanobenzaldehyde sequentially withone equivalent amount of 2-methyl-2-aminoethanol, sodium borohydride,and 2-(2-bromo-ethoxy)-tetrahydro-pyran to obtain a substitutedaminomethylbenzonitrile. Upon removal the tetrahydropyranyl protectiongroup, the benzonitrile can be sequentially mesylated and treated with2-aminomethylpyridine. After the resultant secondary amine is protected,the benzonitrile can then be treated with diisobutylaluminum, oneequivalent amount of a corresponding amine, sodium borohydride, triflicacid, and hydrochloric acid to obtain compounds 114, 118, and 119.

A polyamine compound thus synthesized can be further purified by amethod such as column chromatography, high-pressure liquidchromatography, or recrystallization.

Other polyamine compounds can be prepared using other suitable startingmaterials through the above synthetic routes and others known in theart. The methods described above may also additionally include steps,either before or after the steps described specifically herein, to addor remove suitable protecting groups in order to ultimately allowsynthesis of the polyamine compounds. In addition, various syntheticsteps may be performed in an alternate sequence or order to give thedesired compounds. Synthetic chemistry transformations and protectinggroup methodologies (protection and deprotection) useful in synthesizingapplicable polyamine compounds are known in the art and include, forexample, those described in R. Larock, Comprehensive OrganicTransformations, VCH Publishers (1989); T. W. Greene and P. G. M. Wuts,Protective Groups in Organic Synthesis, 2^(nd) Ed., John Wiley and Sons(1991); L. Fieser and M. Fieser, Fieser and Fieser's Reagents forOrganic Synthesis, John Wiley and Sons (1994); and L. Paquette, ed.,Encyclopedia of Reagents for Organic Synthesis, John Wiley and Sons(1995) and subsequent editions thereof.

The polyamine compounds mentioned herein may contain a non-aromaticdouble bond and one or more asymmetric centers. Thus, they can occur asracemates and racemic mixtures, single enantiomers, individualdiastereomers, diastereomeric mixtures, and cis- or trans-isomericforms. All such isomeric forms are contemplated.

Also within the scope of this invention is a pharmaceutical compositioncontains an effective amount of at least one polyamine compounddescribed above and a pharmaceutical acceptable carrier. Further, thisinvention covers a method of administering an effective amount of one ormore of the polyamine compounds to a patient having a disease describedin the summary section above. This invention also covers a method ofadministering an effective amount of one or more of the polyaminecompounds for enhancing migration of bone marrow-derived cells to blood.“An effective amount” refers to the amount of an active polyaminecompound that is required to confer a therapeutic effect on the treatedsubject. Effective doses will vary, as recognized by those skilled inthe art, depending on the types of diseases treated, route ofadministration, excipient usage, and the possibility of co-usage withother therapeutic treatment.

To practice the method of the present invention, a composition havingone or more polyamine compounds can be administered parenterally,orally, nasally, rectally, topically, or buccally. The term “parenteral”as used herein refers to subcutaneous, intracutaneous, intravenous,intrmuscular, intraarticular, intraarterial, intrasynovial,intrasternal, intrathecal, intralesional, or intracranial injection, aswell as any suitable infusion technique.

A sterile injectable composition can be a solution or suspension in anon-toxic parenterally acceptable diluent or solvent, such as a solutionin 1,3-butanediol. Among the acceptable vehicles and solvents that canbe employed are mannitol, water, Ringer's solution, and isotonic sodiumchloride solution. In addition, fixed oils are conventionally employedas a solvent or suspending medium (e.g., synthetic mono- ordiglycerides). Fatty acid, such as oleic acid and its glyceridederivatives are useful in the preparation of injectables, as are naturalpharmaceutically acceptable oils, such as olive oil or castor oil,especially in their polyoxyethylated versions. These oil solutions orsuspensions can also contain a long chain alcohol diluent or dispersant,carboxymethyl cellulose, or similar dispersing agents. Other commonlyused surfactants such as Tweens or Spans or other similar emulsifyingagents or bioavailability enhancers which are commonly used in themanufacture of pharmaceutically acceptable solid, liquid, or otherdosage forms can also be used for the purpose of formulation.

A composition for oral administration can be any orally acceptabledosage form including capsules, tablets, emulsions and aqueoussuspensions, dispersions, and solutions. In the case of tablets,commonly used carriers include lactose and corn starch. Lubricatingagents, such as magnesium stearate, are also typically added. For oraladministration in a capsule form, useful diluents include lactose anddried corn starch. When aqueous suspensions or emulsions areadministered orally, the active ingredient can be suspended or dissolvedin an oily phase combined with emulsifying or suspending agents. Ifdesired, certain sweetening, flavoring, or coloring agents can be added.

A nasal aerosol or inhalation composition can be prepared according totechniques well known in the art of pharmaceutical formulation. Forexample, such a composition can be prepared as a solution in saline,employing benzyl alcohol or other suitable preservatives, absorptionpromoters to enhance bioavailability, fluorocarbons, and/or othersolubilizing or dispersing agents known in the art.

A composition having one or more active polyamine compounds can also beadministered in the form of suppositories for rectal administration.

The carrier in the pharmaceutical composition must be “acceptable” inthe sense that it is compatible with the active ingredient of thecomposition (and preferably, capable of stabilizing the activeingredient) and not deleterious to the subject to be treated. One ormore solubilizing agents can be utilized as pharmaceutical excipientsfor delivery of an active polyamine compound. Examples of other carriersinclude colloidal silicon oxide, magnesium stearate, cellulose, sodiumlauryl sulfate, and D&C Yellow #10.

The polyamine compounds described above can be preliminarily screenedfor their efficacy in treating above-described diseases by an in vitroassay (See Example 127 below) and then confirmed by animal experimentsand clinic trials. Other methods will also be apparent to those ofordinary skill in the art.

The specific examples below are to be construed as merely illustrative,and not limitative of the remainder of the disclosure in any waywhatsoever. Without further elaboration, it is believed that one skilledin the art can, based on the description herein, utilize the presentinvention to its fullest extent. All publications cited herein arehereby incorporated by reference in their entirety.

EXAMPLE 1 Preparation of Compound 1:N-(4-fluoro-benzyl)-N′,N′-bis-[2-(4-fluoro-benzylamino)-ethyl]-ethane-1,2-diamine

Tris(2-aminoethyl)amine (0.01 mol) and 4-fluoro-benzaldehyde (0.03 mol)were dissolved in MeOH (50 mL). After stirring at 25° C. for 15 h, NaBH₄(1.90 g, 0.05 mol) was added to the above solution at 0° C. The reactionmixture was stirred for 2 h at 25° C. It was then diluted with CH₂Cl₂(100 mL) and ammonium chloride aqueous solution (10%, 70 mL). Theorganic layer was separated, washed with water (100 mL), dried overMgSO₄ (s), and concentrated under reduced pressure to yield an oilproduct. The crude product was purified using alumina columnchromatography (EtOAc/MeOH=8:2) to afford compound 1.

LC/MS (M⁺+1): 471.

EXAMPLE 2 Preparation of Compound 2:N-(3-trifluoromethyl-benzyl)-N′,N′-bis-[2-(3-trifluoromethyl-benzylamino)-ethyl]-ethane-1,2-diamine

Compound 2 was prepared in a manner similar to that described in Example1.

LC/MS (M⁺+1): 621.

EXAMPLE 3 Preparation of Compound 3:N-(3,4-difluoro-benzyl)-N′,N′-bis-[2-(3,4-difluoro-benzylamino)-ethyl]-ethane-1,2-diamine

Compound 3 was prepared in a manner similar to that described in Example1.

LC/MS (M⁺+1): 525.

EXAMPLE 4 Preparation of Compound 4:N-benzyl-N′,N′-bis-(2-benzylamino-ethyl)-ethane-1,2-diamine

Compound 4 was prepared in a manner similar to that described in Example1.

LC/MS (M⁺+1): 417.

EXAMPLE 5 Preparation of Compound 5:N-(2-chloro-4-fluoro-benzyl)-N′,N′-bis-[2-(2-chloro-4-fluoro-benzylamino)-ethyl]-ethane-1,2-diamine

Compound 5 was prepared in a manner similar to that described in Example1.

LC/MS (M⁺+1): 574.

EXAMPLE 6 Preparation of Compound 6:N-(2-fluoro-benzyl)-N′,N′-bis-[2-(2-fluoro-benzylamino)-ethyl]-ethane-1,2-diamine

Compound 6 was prepared in a manner similar to that described in Example1.

LC/MS (M⁺+1): 471.

EXAMPLE 7 Preparation of Compound 7:N-(5-methyl-thiophen-2-ylmethyl)-N′,N′-bis-{2-[(5-methyl-thiophen-2-ylmethyl)-amino]-ethyl}-ethane-1,2-diamine

Compound 7 was prepared in a manner similar to that described in Example1.

LC/MS (M⁺+1): 477.

EXAMPLE 8 Preparation of Compound 8:N-naphthalen-1-ylmethyl-N′,N′-bis-{2-[(naphthalen-1-ylmethyl)-amino]-ethyl}-ethane-1,2-diamine

Compound 8 was prepared in a manner similar to that described in Example1.

LC/MS (M⁺+1): 567.

EXAMPLE 9 Preparation of Compound 9:N-(2,3-dichloro-benzyl)-N′,N′-bis-[2-(2,3-dichloro-benzylamino)-ethyl]-ethane-1,2-diamine

Compound 9 was prepared in a manner similar to that described in Example1.

LC/MS (M⁺+1): 624.

EXAMPLE 10 Preparation of Compound 10:N-(1H-indol-6-ylmethyl)-N′,N′-bis-{2-[(1H-indol-6-ylmethyl)-amino]-ethyl}-ethane-1,2-diamine

Compound 10 was prepared in a manner similar to that described inExample 1.

LC/MS (M⁺+1): 534.

EXAMPLE 11 Preparation of Compound 11:N-(1-methyl-1H-pyrrol-2-ylmethyl)-N′,N′-bis-{2-[(1-methyl-1H-pyrrol-2-ylmethyl)-amino]-ethyl}-ethane-1,2-diamine

Compound 11 was prepared in a manner similar to that described inExample 1.

LC/MS (M⁺+1): 426.

EXAMPLE 12 Preparation of Compound 12:N-[4-({bis-[2-(2-fluoro-benzylamino)-ethyl]-amino}-methyl)-benzyl]-N′-(2-fluoro-benzyl)-N-[2-(2-fluoro-benzylamino)-ethyl]-ethane-1,2-diamine

1,4-Dibromoxylene (0.012 mol) was treated withbis(tert-butoxyaminoethyl)amine (0.024 mol) in the presence of K₂CO₃(0.5 mol) in CH₃CN (60 mL) at 60° C. After stirring for 12 h, thesolution was allowed to cool down to room temperature, filtered, andconcentrated. The concentrate was then treated with HCl/ether andneutralized with K₂CO₃ to afford1,4-di[bis(2-aminoethyl)amino-methyl]benzene quantitatively.

1,4-Di[bis(2-aminoethyl)amino-methyl]benzene (0.01 mol) thus obtainedand 4-fluoro-benzaldehyde (0.04 mol) were dissolved in MeOH (50 mL).After stirring at 25° C. for 15 h, NaBH₄ (2.28 g, 0.06 mol) was added at0° C. to the above solution. The reaction mixture was stirred foranother 2 h at 25° C. It was then diluted with CH₂Cl₂ (100 mL) andammonium chloride aqueous solution (10%, 70 mL). The organic layer wasseparated, washed with water (100 mL), dried over MgSO₄ (s), andconcentrated under reduced pressure to yield an oil product. The crudeproduct was purified using alumina column chromatography(EtOAc/MeOH=7:3) to afford compound 12.

LC/MS (M⁺+1): 741.

EXAMPLE 13 Preparation of Compound 13:N-[4-({bis-[2-(4-fluoro-benzylamino)-ethyl]-amino}-methyl)-benzyl]-N′-(4-fluoro-benzyl)-N-[2-(4-fluoro-benzylamino)-ethyl]-ethane-1,2-diamine

Compound 13 was prepared in a manner similar to that described inExample 12.

LC/MS (M⁺+1): 741.

EXAMPLE 14 Preparation of Compound 14:N-{4-[(bis-{2-[(pyridin-2-ylmethyl)-amino]-ethyl}-amino)-methyl]-benzyl}-N′-pyridin-2-ylmethyl-N-{2-[(pyridin-2-ylmethyl)-amino]-ethyl}-ethane-1,2-diamine

Compound 14 was prepared in a manner similar to that described inExample 12.

LC/MS (M⁺+1): 673.

EXAMPLE 15 Preparation of Compound 15:N-(2-amino-ethyl)-N-{4-[((2-amino-ethyl)-{2-[(pyridin-2-ylmethyl)-amino]-ethyl}-amino)-methyl]-benzyl}-N′-pyridin-2-ylmethyl-ethane-1,2-diamine

1,4-Di[bis(2-aminoethyl)amino-methyl]benzene (0.01 mol) andpyridine-2-carbaldehyde (0.02 mol) were dissolved in MeOH (40 mL). Afterstirring at 25° C. for 15 h, NaBH₄ (1.14 g, 0.03 mol) was added to thissolution at 0° C. The reaction mixture was stirred for another 2 h at25° C. It was then diluted with CH₂Cl₂ (100 mL) and an ammonium chlorideaqueous solution (10%, 70 mL). The organic layer was separated, washedwith water (100 mL), dried over MgSO₄ (s), and concentrated underreduced pressure to yield an oil product. The crude product was purifiedusing alumina column chromatography (EtOAc/MeOH=6:4) to afford compound15.

LC/MS (M⁺+1): 491.

EXAMPLE 16 Preparation of Compound 16:N-(4-fluoro-benzyl)-N′-[2-(4-fluoro-benzylamino)-ethyl]-N′-{4-[(4-fluoro-benzylamino)-methyl]-benzyl}-ethane-1,2-diamine

Bis(2-tert-butoxycarbonylaminoethyl)amine (0.01 mol),4-cyanobenzylbromide (0.01 mol), and K₂CO₃ (0.05 mol) in CH₃CN (70 mL)were heated at 60° C. for 10 h. The resultantbis(2-tert-butoxycarbonylaminoethyl)amino-4-methylphenylcyanide wasdeprotected with HCl/ether and condensed with the 4-fluoro-benzaldehyde(0.02 mol) in MeOH. After sequential treatments with NaBH₄,di-tert-butyl dicarbonate, and diisobutylaluminum,bis(2-substituted-aminoethyl)amino-4-methylbenzaldehyde was obtained andfurther condensed with 4-fluoro-benzylamine to give a Schiff base. TheSchiff base was then reduced by NaBH₄ and deprotected by reacting withHCl. A crude product was obtained and purified with alumina columnchromatography (EtOAc/MeOH=7:3) to afford compound 16.

LC/MS (M⁺+1): 747.

EXAMPLE 17 Preparation of Compound 17:N-{4-[(3-imidazol-1-yl-propylamino)-methyl]-benzyl}-N′-pyridin-2-ylmethyl-N-{2-[(pyridin-2-ylmethyl)-amino]-ethyl}-ethane-1,2-diamine

Compound 17 was prepared in a manner similar to that described inExample 16.

LC/MS (M⁺+1): 513.

EXAMPLE 18 Preparation of Compound 18:1-({4-[(bis-{2-[(pyridin-2-ylmethyl)-amino]-ethyl}-amino)-methyl]-benzylamino}-methyl)-cyclohexanol

Compound 18 was prepared in a manner similar to that described inExample 16.

LC/MS (M⁺+1): 517.

EXAMPLE 19 Preparation of Compound 19:N-{4-[(3-morpholin-4-yl-propylamino)-methyl]-benzyl}-N′-pyridin-2-ylmethyl-N-{2-[(pyridin-2-ylmethyl)-amino]-ethyl}-ethane-1,2-diamine

Compound 19 was prepared in a manner similar to that described inExample 16.

LC/MS (M⁺+1): 532.

EXAMPLE 20 Preparation of Compound 20:N-(4-{[2-(2,5-dimethoxy-phenyl)-ethylamino]methyl}-benzyl)-N′-pyridin-2-ylmethyl-N-{2-[(pyridin-2-ylmethyl)-amino]-ethyl}-ethane-1,2-diamine

Compound 20 was prepared in a manner similar to that described inExample 16.

LC/MS (M⁺+1): 569.

EXAMPLE 21 Preparation of Compound 21:N-(4-{[2-(4-fluoro-phenyl)-1,1-dimethyl-ethylamino]-methyl}-benzyl)-N′-pyridin-2-ylmethyl-N-{2-[(pyridin-2-ylmethyl)-amino]-ethyl}-ethane-1,2-diamine

Compound 21 was prepared in a manner similar to that described inExample 16.

LC/MS (M⁺+1): 555.

EXAMPLE 22 Preparation of Compound 22:N-(4-{[2-(3-fluoro-phenyl)-ethylamino]-methyl}-benzyl)-N′-pyridin-2-ylmethyl-N-{2-[(pyridin-2-ylmethyl)-amino]-ethyl}-ethane-1,2-diamine

Compound 21 was prepared in a manner similar to that described inExample 16.

LC/MS (M⁺+1): 527.

EXAMPLE 23 Preparation of Compound 23:N-(4-{[(1H-benzoimidazol-2-ylmethyl)-amino]-methyl}-benzyl)-N′-pyridin-2-ylmethyl-N-{2-[(pyridin-2-ylmethyl)-amino]-ethyl}-ethane-1,2-diamine

Compound 23 was prepared in a manner similar to that described inExample 16.

LC/MS (M⁺+1): 535.

EXAMPLE 24 Preparation of Compound 24:N-pyridin-2-ylmethyl-N′-{2-[(pyridin-2-ylmethyl)-amino]-ethyl}-N′-(4-{[(pyridin-2-ylmethyl)-amino]-methyl}-benzyl)-ethane-1,2-diamine

Compound 24 was prepared in a manner similar to that described inExample 16.

LC/MS (M⁺+1): 496.

EXAMPLE 25 Preparation of Compound 25:N-(4-{[(pyridin-2-ylmethyl)-amino]-methyl}-benzyl)-N′-(2,3,5-trichloro-benzyl)-N-[2-(2,3,5-trichloro-benzylamino)-ethyl]-ethane-1,2-diamine

Compound 25 was prepared in a manner similar to that described inExample 16.

LC/MS (M⁺+1): 488.

EXAMPLE 26 Preparation of Compound 26:N-(3,4-difluoro-benzyl)-N′-[2-(3,4-difluoro-benzylamino)-ethyl]-N′-(4-{[(pyridin-2-ylmethyl)-amino]-methyl}-benzyl)-ethane-1,2-diamine

Compound 26 was prepared in a manner similar to that described inExample 16.

LC/MS (M⁺+1): 566.

EXAMPLE 27 Preparation of Compound 27:N-(4-fluoro-benzyl)-N′-[2-(4-fluoro-benzylamino)-ethyl]-N′-(4-{[(pyridin-2-ylmethyl)-amino]-methyl}-benzyl)-ethane-1,2-diamine

Compound 27 was prepared in a manner similar to that described inExample 16.

LC/MS (M⁺+1): 530.

EXAMPLE 28 Preparation of Compound 28:N-(4-chloro-benzyl)-N′-[2-(4-chloro-benzylamino)-ethyl]-N′-(4-{[(pyridin-2-ylmethyl)-amino]-methyl}-benzyl)-ethane-1,2-diamine

Compound 28 was prepared in a manner similar to that described inExample 16.

LC/MS (M⁺+1): 563.

EXAMPLE 29 Preparation of Compound 29:N-(4-{[2-(3-chloro-phenyl)-ethylamino]-methyl}-benzyl)-N′-pyridin-2-ylmethyl-N-{2-[(pyridin-2-ylmethyl)-amino]-ethyl}-ethane-1,2-diamine

Compound 29 was prepared in a manner similar to that described inExample 16.

LC/MS (M⁺+1): 543.

EXAMPLE 30 Preparation of Compound 30:N-(4-{[2-(4-chloro-phenyl)-ethylamino]-methyl}-benzyl)-N′-pyridin-2-ylmethyl-N-{2-[(pyridin-2-ylmethyl)-amino]-ethyl}-ethane-1,2-diamine

Compound 30 was prepared in a manner similar to that described inExample 16.

LC/MS (M⁺+1): 543.

EXAMPLE 31 Preparation of Compound 31:N-{4-[(4-fluoro-benzylamino)-methyl]-benzyl}-N′-pyridin-2-ylmethyl-N-{2-[(pyridin-2-ylmethyl)-amino]-ethyl}-ethane-1,2-diamine

Compound 31 was prepared in a manner similar to that described inExample 16.

LC/MS (M⁺+1): 513.

EXAMPLE 32 Preparation of Compound 32:N-(4-{[2-(1H-indol-3-yl)-ethylamino]-methyl}-benzyl)-N′-pyridin-2-ylmethyl-N-{2-[(pyridin-2-ylmethyl)-amino]-ethyl}-ethane-1,2-diamine

Compound 32 was prepared in a manner similar to that described inExample 16.

LC/MS (M⁺+1): 548.

EXAMPLE 33 Preparation of Compound 33:N-(4-{[2-(5-fluoro-1H-indol-3-yl)-ethylamino]-methyl}-benzyl)-N′-pyridin-2-ylmethyl-N-{2-[(pyridin-2-ylmethyl)-amino]-ethyl}-ethane-1,2-diamine

Compound 33 was prepared in a manner similar to that described inExample 16.

LC/MS (M⁺+1): 566.

EXAMPLE 34 Preparation of Compound 34:N-(4-{[2-(5-methoxy-1H-indol-3-yl)-ethylamino]-methyl}-benzyl)-N′-pyridin-2-ylmethyl-N-{2-[(pyridin-2-ylmethyl)-amino]-ethyl}-ethane-1,2-diamine

Compound 34 was prepared in a manner similar to that described inExample 16.

LC/MS (M⁺+1): 578.

EXAMPLE 35 Preparation of Compound 35:N-(4-{[2-(6-methoxy-1H-indol-3-yl)-ethylamino]-methyl}-benzyl)-N′-pyridin-2-ylmethyl-N-{2-[(pyridin-2-ylmethyl)-amino]-ethyl}-ethane-1,2-diamine

Compound 35 was prepared in a manner similar to that described inExample 16.

LC/MS (M⁺+1): 578.

EXAMPLE 36 Preparation of Compound 36:N-(4-{[2-(7-methyl-1H-indol-3-yl)-ethylamino]-methyl}-benzyl)-N′-pyridin-2-ylmethyl-N-{2-[(pyridin-2-ylmethyl)-amino]-ethyl}-ethane-1,2-diamine

Compound 36 was prepared in a manner similar to that described inExample 16.

LC/MS (M⁺+1): 562.

EXAMPLE 37 Preparation of Compound 37:N-{4-[(2-cyclohex-1-enyl-ethylamino)-methyl]-benzyl}-N′-pyridin-2-ylmethyl-N-{2-[(pyridin-2-ylmethyl)-amino]-ethyl}-ethane-1,2-diamine

Compound 37 was prepared in a manner similar to that described inExample 16.

LC/MS (M⁺+1): 513.

EXAMPLE 38 Preparation of Compound 38:N-{4-[(1H-indol-5-ylamino)-methyl]-benzyl}-N′-pyridin-2-ylmethyl-N-{2-[(pyridin-2-ylmethyl)-amino]-ethyl}-ethane-1,2-diamine

Compound 38 was prepared in a manner similar to that described inExample 16.

LC/MS (M⁺+1): 520.

EXAMPLE 39 Preparation of Compound 39:2-{4-[(bis-{2-[(pyridin-2-ylmethyl)-amino]-ethyl}-amino)-methyl]-benzylamino}-4,5,6,7-tetrahydro-benzo[b]thiophene-3-carboxylicacid ethyl ester

Compound 39 was prepared in a manner similar to that described inExample 16.

LC/MS (M⁺+1): 613.

EXAMPLE 40 Preparation of Compound 40:N-(4-{[2-(4-fluoro-phenyl)-ethylamino]-methyl}-benzyl)-N′-pyridin-2-ylmethyl-N-{2-[(pyridin-2-ylmethyl)-amino]-ethyl}-ethane-1,2-diamine

Compound 40 was prepared in a manner similar to that described inExample 16.

LC/MS (M⁺+1): 527.

EXAMPLE 41 Preparation of Compound 41:N-(4-{[2-(4-chloro-phenyl)-propylamino]-methyl}-benzyl)-N′-pyridin-2-ylmethyl-N-{2-[(pyridin-2-ylmethyl)-amino]-ethyl}-ethane-1,2-diamine

Compound 41 was prepared in a manner similar to that described inExample 16.

LC/MS (M⁺+1): 558.

EXAMPLE 42 Preparation of Compound 42:N-(4-{[2-(5-methyl-1H-indol-3-yl)-ethylamino]-methyl}-benzyl)-N′-pyridin-2-ylmethyl-N-{2-[(pyridin-2-ylmethyl)-amino]-ethyl}-ethane-1,2-diamine

Compound 42 was prepared in a manner similar to that described inExample 16.

LC/MS (M⁺+1): 562.

EXAMPLE 43 Preparation of Compound 43:N-(4-fluoro-benzyl)-N′-[2-(3-fluoro-benzylamino)-ethyl]-N′-(4-{[6-(pyridin-2-yloxy)-pyridin-3-ylamino]-methyl}-benzyl)-ethane-1,2-diamine

Compound 43 was prepared in a manner similar to that described inExample 16.

LC/MS (M⁺+1): 609.

EXAMPLE 44 Preparation of Compound 44:6-(5-{4-[(bis-{2-[(pyridin-2-ylmethyl)-amino]-ethyl}-amino)-methyl]-benzylamino}-pyridin-2-yloxy)-pyridine-2-carboxylicacid methyl ester

Compound 44 was prepared in a manner similar to that described inExample 16.

LC/MS (M⁺+1): 633.

EXAMPLE 45 Preparation of Compound 45:N-(4-{[6-(5-chloro-pyridin-2-yloxy)-pyridin-3-ylamino]-methyl}-benzyl)-N′-pyridin-2-ylmethyl-N-{2-[(pyridin-2-ylmethyl)-amino]-ethyl}-ethane-1,2-diamine

Compound 45 was prepared in a manner similar to that described inExample 16.

LC/MS (M⁺+1): 609.

EXAMPLE 46 Preparation of Compound 46:N-pyridin-2-ylmethyl-N′-{2-[(pyridin-2-ylmethyl)-amino]-ethyl}-N′-(4-{[6-(pyridin-2-yloxy)-pyridin-3-ylamino]-methyl}-benzyl)-ethane-1,2-diamine

Compound 46 was prepared in a manner similar to that described inExample 16.

LC/MS (M⁺+1): 575.

EXAMPLE 47 Preparation of Compound 47:N-(4-{[(1H-benzoimidazol-2-ylmethyl)-amino]-methyl}-benzyl)-N′-(4-fluoro-benzyl)-N-[2-(4-fluoro-benzylamino)-ethyl]-ethane-1,2-diamine

Compound 47 was prepared in a manner similar to that described inExample 16.

LC/MS (M⁺+1): 569.

EXAMPLE 48 Preparation of Compound 48:6-{5-[4-({bis-[2-(4-fluoro-benzylamino)-ethyl]-amino}-methyl)-benzylamino]-pyridin-2-yloxy}-pyridine-2-carboxylicacid methyl ester

Compound 48 was prepared in a manner similar to that described inExample 16.

LC/MS (M⁺+1): 667.

EXAMPLE 49 Preparation of Compound 49:N-pyridin-2-ylmethyl-N′-{2-[(pyridin-2-ylmethyl)-amino]-ethyl}-N′-[4-(1,4,8,11tetraaza-cyclotetradec-1-ylmethyl)-benzyl]-ethane-1,2-diamine

K₂CO₃ (0.05 mol) was added to a solution of tri-Boc-protected cyclam(0.01 mole) and 1,4 dibromomethylbenzene (0.01 mol) in CH₃CN at 60° C.After stirring the reaction mixture for 12 h, tri-Boc-protectedbromomethylbenzylcyclam was obtained (0.007 mol). It was then reactedwith bis(2-pyridyliminoethyl)amine (0.01 mole) in CH₃CN (100 mL) in thepresence of K₂CO₃ (0.05 mol) at 60° C. After stirring for 12 h, thereaction mixture was filtered and concentrated. MeOH (50 mL) was addedto this mixture, followed by the addition of NaBH₄ (0.03 mol) at 25° C.The mixture was stirred for another 2 h. The solution was partitionedbetween EtOAc and water. The organic layer was then separated, driedover MgSO₄ (s), filtered, and concentrated to give a residue. Theresidue was treated with HCl/ether and purified by alumina columnchromatography (EtOAc/MeOH=1:2) to afford compound 49.

LC/MS (M⁺+1): 588.

EXAMPLE 50 Preparation of Compound 50:4-[(bis-{2-[(pyridin-2-ylmethyl)-amino]-ethyl}-amino)-methyl]-N,N-bis-{2-[(pyridin-2-ylmethyl)-amino]-ethyl}-benzenesulfonamide

4-Bromomethylbenzenesulfonyl chloride (0.01 mol) was treated withbis(2-tert-butoxycarbonylaminoethyl)amine (0.02 mol) in CH₃CN (100 mL)in the presence of K₂CO₃ (0.1 mol) at 60° C. After stirring for 12 h,the solution was filtered and the filtrate was concentrated to give aresidue. The residue was then treated with HCl/ether and neutralized togive a polyamine. This polyamine was treated with thepyridine-2-carbaldehyde to give a Schiff base. The Schiff base was thenreduced by NaBH₄ in MeOH. The crude product thus obtained was purifiedby alumina column chromatography (EtOAc/MeOH=1:1) to afford compound 50.

LC/MS (M⁺+1): 723.

EXAMPLE 51 Preparation of Compound 51:4-({bis-[2-(3,4-dichloro-benzylamino)-ethyl]-amino}-methyl)-N-pyridin-2-ylmethyl-benzenesulfonamide

4-Bromomethylbenzenesulfonyl chloride (0.01 mol) and 2-aminomethylpyridine (0.01 mol) were dissolved in ether (100 mL), which containsEt₃N (0.02 mol). After stirring for 5 h at 25° C., the solution waswashed with water. The resultant bromosulfamide (0.01 mol) was treatedwith bis(2-tert-butoxycarbonylaminoethyl)amine (0.01 mol) in CH₃CN (100mL) in the presence of K₂CO₃ (0.05 mol) at 60° C. After stirring for 12h, the reaction mixture was filtered and the filtrate was concentratedto give a residue. This residue was treated with HCl/ether andneutralized to give a polyamine. The polyamine was then treated with3,4-dichlorobenzaldehyde to give a Schiff base. The Schiff base wasreduced by NaBH₄ in MeOH. The crude product thus obtained was purifiedby alumina column chromatography (EtOAc/MeOH=6:4) to afford compound 51.

LC/MS (M⁺+1): 680.

EXAMPLE 52 Preparation of Compound 52:4-({bis-[2-(3,4-difluoro-benzylamino)-ethyl]-amino}-methyl)-N-pyridin-2-ylmethyl-benzenesulfonamide

Compound 52 was prepared in a manner similar to that described inExample 51.

LC/MS (M⁺+1): 616.

EXAMPLE 53 Preparation of Compound 53:4-({bis-[2-(4-fluoro-benzylamino)-ethyl]-amino}-methyl)-N-pyridin-2-ylmethyl-benzenesulfonamide

Compound 53 was prepared in a manner similar to that described inExample 51.

LC/MS (M⁺+1): 580.

EXAMPLE 54 Preparation of Compound 54:4-({bis-[2-(4-chloro-benzylamino)-ethyl]-amino}-methyl)-N-pyridin-2-ylmethyl-benzenesulfonamide

Compound 54 was prepared in a manner similar to that described inExample 51.

LC/MS (M⁺+1): 612.

EXAMPLE 55 Preparation of compound 55:4-({bis-[2-(2-chloro-benzylamino)-ethyl]-amino}-methyl)-N-pyridin-2-ylmethyl-benzenesulfonamide

Compound 55 was prepared in a manner similar to that described inExample 51.

LC/MS (M⁺+1): 612.

EXAMPLE 56 Preparation of Compound 56:N-[4-(6-methoxy-1,3,4,9-tetrahydro-b-carbolin-2-ylmethyl)-benzyl]-N′-pyridin-2-ylmethyl-N-{2-[(pyridin-2-ylmethyl)-amino]-ethyl}-ethane-1,2-diamine

1,4-Dibromoxylene (13.64 mmol) was treated withbis(tert-butoxyaminoethyl)amine (2.74 mmol) in the presence of Et₃N(2.74 mmol) in CH₂Cl₂ (50 mL) at 0° C. After stirring 16 h, the solutionwas filtered, concentrated, and purified to afford a mono-substitutedbromide. This mono-substituted bromide (0.68 mmol) was reacted with6-methoxy-1,2,3,4-tetrahydro-9H-pyrido[3,4-b]indole (0.68 mmol) in CH₃CN(10 mL) in the presence of K₂CO₃ (3.39 mmol) at 60° C. After stirringfor 12 h, the solution was filtered and the filtrate was concentratedand purified by chromatography to give a Boc-protected residue (0.57mmol; 84% yield). This residue (0.26 mmol) was treated with HCl/etherand neutralized to give a polyamine. This polyamine was then treatedwith the pyridine-2-carbaldehyde to give a Schiff base. The Schiff basewas reduced by NaBH₄ in MeOH. The crude product thus obtained waspurified by alumina column chromatography (EtOAc/MeOH=7:3) to affordcompound 56.

LC/MS (M⁺+1): 590.

EXAMPLE 57 Preparation of Compound 57:N-pyridin-2-ylmethyl-N′-{2-[(pyridin-2-ylmethyl)-amino]-ethyl}-N′-[4-(1,3,4,9-tetrahydro-b-carbolin-2-ylmethyl)-benzyl]-ethane-1,2-diamine

Compound 57 was prepared in a manner similar to that described inExample 56.

LC/MS (M⁺+1): 560.

EXAMPLE 58 Preparation of Compound 58: isonicotinic acid{4-[(bis-{2-[(pyridin-2-ylmethyl)-amino]-ethyl}-amino)-methyl]-benzylidene}-hydrazide

Bis(2-tert-butoxycarbonylaminoethyl)amine (0.01 mol),4-cyanobenzylbromide (0.01 mol), and K₂CO₃ (0.05 mol) were added inCH₃CN (70 mL) and the mixture were heated at 60° C. while stirring for10 h. The resultantbis(2-tert-butoxycarbonylaminoethyl)amino-4-methylphenylcyanide wasdeprotected by treating with HCl/ether and then condensed withpyridine-2-benzaldehyde (0.02 mol) in MeOH. After sequential treatmentswith NaBH₄, di-tert-butyl dicarbonate, and DIBAL, the resultantbis(2-substituted-aminoethyl)amino-4-methylbenzaldehyde was condensedwith isonicotinic acid hydrazide to give a Schiff base. The Schiff basewas then treated with HCl/ether. The crude product thus obtained waspurified by alumina column chromatography (EtOAc/MeOH=7:3) to affordcompound 58.

LC/MS (M⁺+1): 523.

EXAMPLE 59 Preparation of Compound 59: thiophene-2-carboxylic acid{4-[(bis-{2-[(pyridin-2-ylmethyl)-amino]-ethyl}-amino)-methyl]-benzylidene}-hydrazide

Compound 59 was prepared in a manner similar to that described inExample 58.

LC/MS (M⁺+1): 528.

EXAMPLE 60 Preparation of Compound 60:1-[bis[(2-(2-pyridinylmethyl)aminoethyl)]amino-methyl]-4-[2-(3-indol)-1-((S)-hydroxymethyl)ethylaminomethyl]benzene

Compound 60 was prepared in a manner similar to that described inExample 16 (Yield: 80%).

LC-MS (C₃₅H₄₃N₇O.7HCl) (M⁺+1-7HCl): 578.

EXAMPLE 61 Preparation of Compound 61:1-[bis[(2-(2-pyridinyl-2-ethyl)aminoethyl)]amino-methyl]-4-[2-(3-indol)-1-((S)-hydroxymethyl)ethylaminomethyl]benzene

Compound 61was prepared in a manner similar to that described in Example16 (Yield: 80%).

LC-MS (C₃₅H₄₃N₇O.7HCl) (M⁺+1-7HCl): 606.

EXAMPLE 62 Preparation of Compound 62:1-[bis[(2-(2-pyridinylmethyl)aminoethyl)]amino-methyl]-4-[2-(phenyl)-1-((R)-hydroxymethyl)ethylaminomethyl]benzene

Compound 62 was prepared in a manner similar to that described inExample 16 (Yield: 81%).

LC-MS (C₃₃H₄₂N₆O.6HCl) (M⁺+1-6HCl): 539.

EXAMPLE 63 Preparation of Compound 63:1-[bis[(2-(2-pyridinylmethyl)aminoethyl)]amino-methyl]-4-[2-(phenyl)-1-((S)-hydroxymethyl)ethylaminomethyl]benzene

Compound 63 was prepared in a manner similar to that described inExample 16 (Yield: 85%).

LC-MS (C₃₃H₄₂N₆O.6HCl) (M⁺+1-6HCl): 539.

EXAMPLE 64 Preparation of Compound 64:1-[bis[(2-(2-pyridinylmethyl)aminoethyl)]amino-methyl]-4-[2-(3-indol)-1-((R)-hydroxymethyl)ethylaminomethyl]benzene

Compound 64 was prepared in a manner similar to that described inExample 16 (Yield: 78%).

LC-MS (C₃₅H₄₃N₇O.7HCl) (M⁺+1-7HCl): 578.

EXAMPLE 65 Preparation of Compound 65:1-[bis[(2-(2-pyridinyl-2-ethyl)aminoethyl)]amino-methyl]-4-[(1,2,3,4-tetrahydro-9H-pyrido-6-methoxy[3,4-b]indol-2-methyl)benzene

Compound 65 was prepared by a similar manner to that described inExample 56.

LC/MS (M⁺+1): 618.

EXAMPLE 66 Preparation of Compound 66:1-[bis[(2-(2-pyridinylmethyl)aminoethyl)]amino-methyl]-4-[(1,2,3,4-tetrahydro-9H-pyrido-6-benzyloxy[3,4-b]indol-2-methyl)benzene

Compound 66 was prepared by a similar manner to that described inExample 56.

LC/MS (M⁺+1): 666.

EXAMPLE 67 Preparation of Compound 67:1-[bis[(2-(2-pyridinylmethyl)aminoethyl)]amino-sulfonyl]-4-(1,2,3,4-tetrahydro-9H-pyrido-6-methoxy[3,4-b]indol-2-methyl)benzene

Bis(2-tert-butoxycarbonylaminoethyl)amine (3.03 g, 0.01 mol) was addedto a solution of 1-bromomethylbenzene-4-sulfonyl chloride (2.68 g, 0.01mol), CH₂Cl₂ (160 mL), and Et₃N (1.01 g, 0.01 mol). The reaction mixturewas stirred at 0° C. for 2.5 hours. Then, the solvent was evaporated andthe residue was dissolved in a mixture of CH₃CN (180 mL), K₂CO₃ (4.14 g,0.03 mol), and 1,2,3,4-tetrahydro-9H-pyrido-6-methoxy[3,4-b]indole (1.72g, 0.01 mol). The reaction mixture was stirred at 60° C. for another 10hours. The mixture was then filtered, concentrated, and treated with amixture of CH₂Cl₂ (35 mL) and HCl/ether (1.0 M, 80 mL) for 12 hours. Thereaction mixture was subsequently concentrated, stirred with anhydrousK₂CO₃ (10.0 g, 30 min) in CH₂Cl₂ (150 mL). The mixture thus obtained wasfiltered and concentrated to afford intermediate 1,1-[bis[(2-aminoethyl)]aminosulfonyl]-4-[(1,2,3,4-tetrahydro-9H-pyrido-6-methoxy[3,4-b]indol-2-methyl)benzene(2.56 g, 0.006 mol) in 60% yield. This intermediate was then treatedwith pyridine-2-carboxaldehyde (1.50 g, 0.014 mol) in MeOH (40 mL) for14 hours and then with NaBH₄ (1.60 g, 0.042 mol) for 4 hours to give acrude intermediate 2. Intermediate 2 was purified using alumina columnchromatography (EtOAc/MeOH=7:3) (3.19 g, 0.005 mol, 83% yield).Intermediate 2 was subsequently treated with HCl/ether (125 mL) inCH₂Cl₂ (50 mL) to afford compound 67.

LC-MS (C₃₅H₄₁N₇O₃S.6HCl) (M⁺+1-6HCl): 640.

EXAMPLE 68 Preparation of Compound 68:N-[4-(6-methoxy-1,3,4,9-tetrahydro-b-carbolin-2-ylmethyl)-benzyl]-N′-pyridin-2-ylmethyl-N-{2-[(pyridin-2-ylmethyl)-2-hydroxyethylamino]-ethyl}-ethane-1,2-diamine

Compound 68 was prepared by selective alkylation of compound 56 (Yield:40%).

LC-MS (C₃₈H₄₇N₇O₂.7HCl) (M⁺+1-7HCl): 634.

EXAMPLE 69 Preparation of Compound 69:1-[bis[(2-(2-pyridinylmethyl)aminoethyl)]amino-methyl]-4-[2-(phenyl)-1-((R)-hydroxycarbonyl)ethylaminomethyl]benzene

Compound 69 was prepared in a manner similar to that described inExample 16 (Yield: 70%).

LC-MS (C₃₃H₄₀N₆O₂.6HCl) (M⁺+1-6HCl): 553.

EXAMPLE 70 Preparation of Compound 70:1-[bis[(2-(2-pyridinylmethyl)aminoethyl)]amino-methyl]-4-[2-(3-(5-hydroxyindol)-1-((R)-hydroxycarbonyl)ethylaminomethyl]benzene

Compound 70 was prepared in a manner similar to that described inExample 16 (Yield: 75%).

LC-MS (C₃₅H₄₁N₇O₃.7HCl) (M⁺+1-7HCl): 608.

EXAMPLE 71 Preparation of Compound 71:1-[bis[(2-(2-pyridinylmethyl)aminoethyl)]amino-methyl]-4-[2-(3-(7-methylindol))ethylaminomethyl]benzene

Compound 71 was prepared in a manner similar to that described inExample 16 (Yield: 70%).

LC-MS (C₃₇H₄₇N₇.7HCl) (M⁺+1-7HCl): 590.

EXAMPLE 72 Preparation of Compound 72:1-[bis[(2-(2-pyridinyl-2-ethyl)aminoethyl)]amino-methyl]-4-[2-(3-chlorophenyl)ethylaminomethyl]benzene

Compound 72 was prepared in a manner similar to that described inExample 16 (Yield: 80%).

LC-MS (C₃₄H₄₃ClN₆.6HCl) (M⁺+1-6HCl): 571.

EXAMPLE 73 Preparation of Compound 73:1-[bis[(2-(2-pyridinyl-2-ethyl)aminoethyl)]amino-methyl]-4-{[(1H-benzoimidazol-2-ylmethyl)-amino]-methyl}benzene

Compound 73 was prepared in a manner similar to that described inExample 16 (Yield: 84%).

LC-MS (C₃₄H₄₂N₈.7HCl) (M⁺+1-6HCl): 563.

EXAMPLE 74 Preparation of Compound 74:1-[bis[(2-(2-pyridinylmethyl)aminoethyl)]amino-methyl]-4-[(2-phenyl-2-hydroxy-1-methoxymethyl)ethylaminomethyl]benzene

Compound 74 was prepared in a manner similar to that described inExample 16 (Yield: 71%).

LC-MS (C₃₄H₄₄N₆O₂.6HCl) (M⁺+1-6HCl): 569.

EXAMPLE 75 Preparation of Compound 75:1-[bis[(2-(2-pyridinylmethyl)aminoethyl)]amino-methyl]-4-[(2-(4-chlorophenyl)-1-hydroxymethyl)ethylaminomethyl]benzene

Compound 75 was prepared in a manner similar to that described inExample 16 (Yield: 70%).

LC-MS (C₃₃H₄₁ClN₆O.6HCl) (M⁺+1-6HCl): 573.

EXAMPLE 76 Preparation of Compound 76:1-[bis[(2-(2-pyridinylmethyl)aminoethyl)]amino-methyl]-4-[(2-phenyl-(2R)-hydroxy-(1R)-hydroxymethyl)ethylaminomethyl]benzene

Compound 76 was prepared in a manner similar to that described inExample 16 (Yield: 68%).

LC-MS (C₃₃H₄₂N₆O₂.6HCl) (M⁺+1-6HCl): 555.

EXAMPLE 77 Preparation of Compound 77:1-[bis[(2-(2-pyridinylmethyl)aminoethyl)]amino-methyl]-4-[(2-phenyl-(2S)-hydroxy-(1S)-hydroxymethyl)ethylaminomethyl]benzene

Compound 77 was prepared in a manner similar to that described inExample 16 (Yield: 70%).

LC-MS (C₃₃H₄₂N₆O₂.6HCl) (M⁺+1-6HCl): 555.

EXAMPLE 78 Preparation of Compound 78:1-[bis[(2-(2-pyridinylmethyl)aminoethyl)]amino-methyl]-4-[2-(4-bromophenyl)-1-((R)-hydroxycarbonyl)ethylaminomethyl]benzene

Compound 78 was prepared in a manner similar to that described inExample 16 (Yield: 77%).

LC-MS (C₃₃H₃₉BrN₆O₂.6HCl) (M⁺+1-6HCl): 631.

EXAMPLE 79 Preparation of Compound 79:2-({4′-[(bis-{2-[(pyridin-2-ylmethyl)-amino]-ethyl}-amino)-methyl]-biphenyl-4-ylmethyl}-pyridin-2-ylmethyl-amino)-3-phenyl-propan-1-ol

Compound 79 was prepared in a manner similar to that described inExample 16 (Yield: 70%).

LC-MS (C₄₅H₅₁N₇O.7HCl) (M⁺+1-7HCl): 706.

EXAMPLE 80 Preparation of Compound 80:N-pyridin-2-ylmethyl-N′-{2-[(pyridin-2-ylmethyl)-amino]-ethyl}-N′-[4-(6-chloro-1,3,4,9-tetrahydro-b-carbolin-2-ylmethyl)-benzyl]-ethane-1,2-diamine

Compound 80 was prepared in a manner similar to that described inExample 56.

LC-MS (C₃₅H₄₀ClN₇.7HCl) (M⁺+1-7HCl): 594.

EXAMPLE 81 Preparation of Compound 81:N-[4-({ethyl-[2-(7-methyl-1H-indol-3-yl)-ethyl]-amino}-methyl)-benzyl]-N′-pyridin-2-ylmethyl-N-{2-[(pyridin-2-ylmethyl)-amino]-ethyl}-ethane-1,2-diamine

Compound 81 was prepared in a manner similar to that described inExample 56.

LC-MS (C₃₇H₄₇N₇.7HCl) (M⁺+1-7HCl): 590.

EXAMPLE 82 Preparation of Compound 82:N-[4-({[2-(3-chloro-phenyl)-ethyl]-ethyl-amino}-methyl)-benzyl]-N′-pyridin-2-ylmethyl-N-{2-[(pyridin-2-ylmethyl)-amino]-ethyl}-ethane-1,2-diamine

Compound 82 was prepared in a manner similar to that described inExample 56.

LC-MS (C₃₄H₄₃ClN₆.6HCl) (M⁺+1-6HCl): 571.

EXAMPLE 83 Preparation of Compound 83:1-[bis[(2-(2-pyridinylmethyl)aminoethyl)]amino-methyl]-4-[(2S)-hydroxymethylpyrrolidine-N-methyl]benzene

Compound 83 was prepared in a manner similar to that described inExample 56 (Yield: 80%).

LC-MS (C₂₉H₄₀N₆O.6HCl) (M⁺+1-6HCl): 489.

EXAMPLE 84 Preparation of Compound 84:1-[bis[(2-(2-pyridinylmethyl)aminoethyl)]amino-methyl]-4-[(2R)-hydroxymethylpyrrolidine-N-methyl]benzene

Compound 84 was prepared in a manner similar to that described inExample 56 (Yield: 80%).

LC-MS (C₂₉H₄₀N₆O.6HCl) (M⁺+1-6HCl): 489.

EXAMPLE 85 Preparation of Compound 85:2-({4′-[(bis-{2-[(pyridin-2-ylmethyl)-amino]-ethyl}-amino)-methyl]-biphenyl-4-ylmethyl}-amino)-3-phenyl-propan-1-ol

Compound 85 was prepared in a manner similar to that described inExample 16.

LC-MS (C₃₉H₄₆N₆O.6HCl) (M⁺+1-6HCl): 615.

EXAMPLE 86 Preparation of Compound 86:N-pyridin-2-ylmethyl-N′-{2-[(pyridin-2-ylmethyl)-amino]-ethyl}-N′-[4-(6-fluoro-1,3,4,9-tetrahydro-b-carbolin-2-ylmethyl)-benzyl]-ethane-1,2-diamine

Compound 86 was prepared in a manner similar to that described inExample 56.

LC-MS (C₃₅H₄₀FN₇.7HCl) (M⁺+1-7HCl): 578.

EXAMPLE 87 Preparation of Compound 87:1-[bis[(2-(2-pyridinylmethyl)aminoethyl)]amino-methyl]-4-[2-(5-chloro-6-hydroxyphenyl-1-hydroxycarbonyl)ethylaminomethyl]benzene

Compound 87 was prepared in a manner similar to that described inExample 16 (Yield: 76%).

LC-MS (C₃₃H₃₉ClN₆O₃.6HCl) (M⁺+1-6HCl): 603.

EXAMPLE 88 Preparation of Compound 88:1-[bis[(2-(2-pyridinylmethyl)aminoethyl)]amino-methyl]-4-[(2-thiophene-1-hydroxycarbonyl)ethylaminomethyl]benzene

Compound 88 was prepared in a manner similar to that described inExample 16 (Yield: 70%).

LC-MS (C₃₁H₃₈ClN₆O₂S.6HCl) (M⁺+1-6HCl): 559.

EXAMPLE 89 Preparation of Compound 89:1-[bis[(2-(2-pyridinylmethyl)aminoethyl)]amino-methyl]-4-{[2-(4-chlorophenyl)-2-cyclopropyl]ethylaminomethyl}benzene

Compound 89 was prepared in a manner similar to that described inExample 16 (Yield: 78%).

LC-MS (C₃₄H₄₁ClN₆.6HCl) (M⁺+1-6HCl): 569.

EXAMPLE 90 Preparation of Compound 90:1-[bis[(2-(2-pyridinylmethyl)aminoethyl)]amino-methyl]-4-{[2-(3-chloro-6-methoxyphenyl)]ethylaminomethyl}benzene

Compound 90 was prepared in a manner similar to that described inExample 16 (Yield: 80%).

LC-MS (C₃₃H₄₁ClN₆O.6HCl) (M⁺+1-6HCl): 573.

EXAMPLE 91 Preparation of Compound 91:N-pyridin-2-yl-2-ethyl-N′-{2-[(pyridin-2-ylmethyl)-amino]-ethyl}-N′-[4-(6-methoxy-1,3,4,9-tetrahydro-b-carbolin-2-ylmethyl)-benzyl]-ethane-1,2-diamine

Compound 91 was prepared in a manner similar to that described inExample 56 (Yield: 65%).

LC-MS (C₃₇H₄₅N₇O.7HCl) (M⁺+1-7HCl): 604.

EXAMPLE 92 Preparation of Compound 92:1-[bis[(2-(2-pyridinylmethyl)aminoethyl)]amino-methyl]-4-[(2-thiophene-2-hydroxycarbonyl)ethylaminomethyl]benzene

Compound 92 was prepared in a manner similar to that described inExample 16 (Yield: 85%).

LC-MS (C₃₁H₃₈N₆O₂S.6HCl) (M⁺+1-6HCl): 559.

EXAMPLE 93 Preparation of Compound 93:1-[bis[(2-(2-pyridinylmethyl)aminoethyl)]amino-methyl]-4-[3-hydroxymethyl-(S)-1,2,3,4-tetrahydroisoquinoline-N-ylmethyl]benzene

Compound 93 was prepared in a manner similar to that described inExample 56.

LC-MS (C₃₄H₄₂N₆O.6HCl) (M⁺+1-6HCl): 551.

EXAMPLE 94 Preparation of Compound 94:1-[bis[(2-(2-pyridinylmethyl)aminoethyl)]amino-methyl]-4-(4-hydroxy-4-phenylpiperidine-N-ylmethyl)benzene

Compound 94 was prepared in a manner similar to that described inExample 56.

LC-MS (C₃₅H₄₄N₆O.6HCl) (M⁺+1-6HCl): 565.

EXAMPLE 95 Preparation of Compound 95:1-[bis[(2-(2-pyridinylmethyl)aminoethyl)]amino-methyl]-4-[4-hydroxy-4-(4-chlorophenylpiperidine)-N-ylmethyl]benzene

Compound 95 was prepared in a manner similar to that described inExample 56.

LC-MS (C₃₅H₄₃ClN₆O.6HCl) (M⁺+1-6HCl): 599.

EXAMPLE 96 Preparation of Compound 96:1-[bis[(2-(2-pyridinylmethyl)aminoethyl)]amino-methyl]-4-(1-indanaminomethyl)benzene

Compound 96 was prepared in a manner similar to that described inExample 16.

LC-MS (C₃₃H₄₀N₆.6HCl) (M⁺+1-6HCl): 521.

EXAMPLE 97 Preparation of Compound 97:1-[bis[(2-(2-pyridinylmethyl)aminoethyl)]amino-methyl]-4-(3,4-dichlorophenylmethylaminomethyl)benzene

Compound 97 was prepared in a manner similar to that described inExample 16.

LC-MS (C₃₁H₃₆Cl₂N₆.6HCl) (M⁺+1-6HCl): 563.

EXAMPLE 98 Preparation of Compound 98:1-[bis[(2-(2-pyridinylmethyl)aminoethyl)]amino-methyl]-4-(3-chlorophenylmethylaminomethyl)benzene

Compound 98 was prepared in a manner similar to that described inExample 16.

LC-MS (C₃₁H₃₇ClN₆.6HCl) (M⁺+1-6HCl): 529.

EXAMPLE 99 Preparation of Compound 99:1-[bis[(2-(2-pyridinylmethyl)aminoethyl)]amino-methyl]-4-[4-hydroxyphenyl(1-hydroxymethyl)ethylaminomethyl)benzene

Compound 99 was prepared in a manner similar to that described inExample 16.

LC-MS (C₃₃H₄₂N₆O₂.6HCl) (M⁺+1-6HCl): 555.

EXAMPLE 100 Preparation of Compound 100:N-(4-{[2-(7-methyl-1H-indol-3-yl)-1-(hydroxycarbonylmethyl)ethylamino]-methyl}-benzyl)-N′-pyridin-2-ylmethyl-N-{2-[(pyridin-2-ylmethyl)-amino]-ethyl}-ethane-1,2-diamine

Compound 100 was prepared in a manner similar to that described inExample 16.

LC-MS (C₃₇H₄₅N₇O₂.6HCl) (M⁺+1-6HCl): 620.

EXAMPLE 101 Preparation of Compound 101:N-(4-{[2-(7-methyl-1H-indol-3-yl)-1-(hydroxyethyl)ethylamino]-methyl}-benzyl)-N′-pyridin-2-ylmethyl-N-{2-[(pyridin-2-ylmethyl)-amino]-ethyl}-ethane-1,2-diamine

Compound 101 was prepared in a manner similar to that described inExample 16.

LC-MS (C₃₇H₄₇N₇O.7HCl) (M⁺+1-7HCl): 606.

EXAMPLE 102 Preparation of Compound 102:1-[bis[(2-(2-pyridinylmethyl)aminoethyl)]amino-methyl]-4-{[2-(3-chloro-4-ethylphenyl)]ethylaminomethyl}benzene

Compound 102 was prepared in a manner similar to that described inExample 16 (Yield: 80%).

LC-MS (C₃₃H₄₁ClN₆O.6HCl) (M⁺+1-6HCl): 573.

EXAMPLE 103 Preparation of Compound 103:N-(4-{[2-(7-methyl-1H-indol-3-yl)ethylamino]-methyl}-benzyl)-N′-pyridin-2-ylmethyl-N-{2-[(pyridin-2-ylmethyl)-amino]-ethyl}-propane-1,3-diamine

Compound 103 was prepared in a manner similar to that described inExample 16.

LC-MS (C₃₆H₄₅N₇.7HCl) (M⁺+1-7HCl): 576.

EXAMPLE 104 Preparation of Compound 104:N-(4-{[2-(3-chlorophenyl)ethylamino]methyl}-benzyl)-N′-pyridin-2-ylmethyl-N-{2-[(pyridin-2-ylmethyl)-amino]-ethyl}-propane-1,3-diamine

Compound 104 was prepared in a manner similar to that described inExample 16.

LC-MS (C₃₃H₄₁ClN₆.6HCl) (M⁺+1-6HCl): 557.

EXAMPLE 105 Preparation of Compound 105:N-(4-{[(1H-benzoimidazol-2-yl-methyl)amino]-methyl}-benzyl)-N′-pyridin-2-ylmethyl-N-{2-[(pyridin-2-ylmethyl)-amino]-ethyl}-propane-1,3-diamine

Compound 105 was prepared in a manner similar to that described inExample 16.

LC-MS (C₃₃H₄₀N₈.7HCl) (M⁺+1-7HCl): 549.

EXAMPLE 106 Preparation of Compound 106:N-pyridin-2-yl-hydroxyethyl-N′-{2-[(pyridin-2-ylmethyl)-amino]-ethyl}-N′-[4-(6-methoxy-1,3,4,9-tetrahydro-b-carbolin-2-ylmethyl)-benzyl]-ethane-1,2-diamine

Compound 106 was prepared in a manner similar to that described inExample 56 (Yield: 65%).

LC-MS (C₃₇H₄₅N₇O₂.7HCl) (M⁺+1-7HCl): 620.

EXAMPLE 107 Preparation of Compound 107:1-[bis[(2-(2-pyridinylmethyl)aminoethyl)]amino-methyl]-4-(2-methoxymethylpyrrolidine-N-methylbenzene

Compound 107 was prepared in a manner similar to that described inExample 56 (Yield: 82%).

LC-MS (C₃₀H₄₂N₆O.6HCl) (M⁺+1-6HCl): 503.

EXAMPLE 108 Preparation of Compound 108:N-pyridin-2-yl-hydroxyethyl-N′-{2-[(pyridin-2-ylmethyl)-amino]-ethyl}-N′-[4-(1-hydroxymethyl-6-methoxy-1,3,4,9-tetrahydro-b-carbolin-2-yl-methyl)-benzyl]-ethane-1,2-diamine

Compound 108 was prepared in a manner similar to that described inExample 56.

LC-MS (C₃₇H₄₅N₇O₂.7HCl) (M⁺+1-7HCl): 620.

EXAMPLE 109 Preparation of Compound 109:N-pyridin-2-yl-hydroxyethyl-N′-{2-[(pyridin-2-ylmethyl)-amino]-ethyl}-N′-[4-(4-phenyl-1,2,3,6-tetrahydropyridine-1-methyl)-benzyl]-ethane-1,2-diamine

Compound 109 was prepared in a manner similar to that described inExample 56.

LC-MS (C₃₅H₄₂N₆.6HCl) (M⁺+1-6HCl): 547.

EXAMPLE 110 Preparation of Compound 110:N-pyridin-2-yl-2-ethyl-N′-{2-[(pyridin-2-yl-ethyl)-amino]-ethyl}-4-(cyanobenzyl]-ethane-1,2-diamine.5HCl

A mixture of bis(2-tert-butoxycarbonylaminoethyl)amine (0.01 mol),4-cyanobenzyl-bromide (0.01 mol), K₂CO₃ (0.05 mol), and CH₃CN (70 mL)was heated at 60° C. for 10 hours. The resultantbis(2-tert-butoxycarbonylaminoethyl)amino-4-methylphenylcyanide wasdeprotected using HCl/ether, condensed with 2-acetyl pyridine (0.01 mol)in MeOH, and then reduced by NaBH₄. After sequential treatments withpyridine-2-carboxaldehyde, NaBH₄, and HCl, compound 110 was obtained in75% overall yield.

LC-MS (C₂₅H₃₀N₆.5HCl) (M⁺+1-5HCl): 415.

EXAMPLE 111 Preparation of Compound 111:N-pyridin-2-yl-2-ethyl-N′-{2-[(pyridin-2-ylmethyl)-amino]-ethyl}-N′-[4-(1H-benzimidazol-2-yl-methyl)-aminomethyl]benzyl]-ethane-1,2-diamine

A mixture of bis(2-tert-butoxycarbonylaminoethyl)amine (0.01 mol),4-cyanobenzyl-bromide (0.01 mol), K₂CO₃ (0.05 mol), and CH₃CN (70 mL)was heated at 60° C. for 10 hours. The resultantbis(2-tert-butoxycarbonylaminoethyl)amino-4-methylphenylcyanide wasdeprotected by HCl/ether, condensed with 2-acetyl pyridine (0.01 mol) inMeOH, and then reduced by NaBH₄. After sequential treatments withpyridine-2-carboxaldehyde, NaBH₄, di-tert-butyl dicarbonate, anddiisobutylaluminum, the aldehyde thus obtained was condensed with2-aminomethylbenzimidazole to give a Schiff base. The Schiff base wasthen reduced by NaBH₄ and deprotected by HCl to afford compound 111 in45% overall yield.

LC-MS (C₃₃H₄₀N₈.7HCl) (M⁺+1-7HCl): 549.

EXAMPLE 112 Preparation of Compound 112:1-[bis[(2-(2-pyridinylmethyl)aminoethyl)]amino-methyl]-4-[4-hydroxy-4-(4-methoxyphenylpiperidine)-N-yl-methyl]benzene

Compound 112 was prepared in a manner similar to that described inExample 67.

LC-MS (C₃₆H₄₆N₆O₂.6HCl) (M⁺+1-6HCl): 595.

EXAMPLE 113 Preparation of Compound 113:1-[bis[(2-(2-pyridinylmethyl)aminoethyl)]amino-methyl]-4-[4-hydroxy-4-(2-methoxyphenylpiperidine)-N-yl-methyl]benzene

Compound 113 was prepared in a manner similar to that described inExample 56.

LC-MS (C₃₆H₄₆N₆O₂.6HCl) (M⁺+1-6HCl): 595.

EXAMPLE 114 Preparation of Compound 114:N-(4-{[2-(7-methyl-1H-indol-3-yl)ethylamino]-methyl}-benzyl)-N′-pyridin-2-ylmethyl-N-{2-[(pyridin-2-ylmethyl)-amino]-ethyl}-1-methylethane-1,2-diamine

4-Cyanobenzaldehyde (0.01 mol) was treated with 2-methyl-2-aminoethanol(0.01 mol) in MeOH (20 mL) at 60° C. for 12 hours. NaBH₄ (1.90 g, 0.05mol) was then added to the above solution at 0° C. The reaction mixturewas stirred for another 2 hours at 25° C. It was then diluted withCH₂Cl₂ (100 mL) and with an ammonium chloride aqueous solution (10%, 70mL). The organic layer was separated, washed with water (100 mL), driedover MgSO₄ (s), and concentrated under reduced pressure to yield an oilintermediate. The oil intermediate was then condensed with2-(2-bromo-ethoxy)-tetrahydro-pyran (0.01 mol) in the presence of K₂CO₃(0.05 mol) in refluxing CH₃CN (30 mL). After deprotection, the resultanthydroxyl group was mesylated and was allowed to react with2-aminomethylpyridine. The resultant secondary amine was then protectedwith the Boc group. Subsequently, the cyanide group was treated withdiisobutylaluminum and the aldehyde thus obtained was sequentiallytreated with 6-methyl-3-aminoethylindol, NaBH₄, triflic acid, and HCl toafford compound 114 in 60% overall yield.

LC-MS (C₃₆H₄₅N₇.6HCl) (M⁺+1-6HCl): 576.

EXAMPLE 115 Preparation of Compound 115:2-{4-[(bis-{2-[(pyridin-2-ylmethyl)-amino]-ethyl}-amino)-methyl]-benzylamino}-1-(3-chloro-phenyl)-ethanol

Compound 115 was prepared in a manner similar to that described inExample 16 (Yield: 77%).

LC-MS (M⁺+1): 559.

EXAMPLE 116 Preparation of Compound 116:N-(4-{[(1H-benzoimidazol-2-ylmethyl)-amino]-methyl}-benzyl)-N′-piperidin-4-yl-N-{2-[(pyridin-2-ylmethyl)-amino]-ethyl}-ethane-1,2-diamine

Compound 116 was prepared in a manner similar to that described inExample 111 (Yield: 70%).

LC-MS (M⁺+1): 527.

EXAMPLE 117 Preparation of Compound 117:N-(4-{[2-(7-methyl-1H-indol-3-yl)-ethylamino]-methyl}-benzyl)-N′-(1-pyridin-2-yl-ethyl)-N-{2-[(pyridin-2-ylmethyl)-amino]-ethyl}-ethane-1,2-diamine

Compound 117 was prepared in a manner similar to that described inExample 111 (Yield: 70%).

LC-MS (M⁺+1): 576.

EXAMPLE 118 Preparation of Compound 118:N2-(4-{[(1H-benzoimidazol-2-ylmethyl)-amino]-methyl}-benzyl)-N1-pyridin-2-ylmethyl-N2-{2-[(pyridin-2-ylmethyl)-amino]-ethyl}-propane-1,2-diamine

Compound 118 was prepared in a manner similar to that described inExample 114 (Yield: 68%).

LC-MS (M⁺+1): 549.

EXAMPLE 119 Preparation of Compound 119:N2-(4-{[2-(3-chloro-phenyl)-ethylamino]-methyl}-benzyl)-N1-pyridin-2-ylmethyl-N2-{2-[(pyridin-2-ylmethyl)-amino]-ethyl}-propane-1,2-diamine

Compound 119 was prepared in a manner similar to that described inExample 114 (Yield: 69%).

LC-MS (M⁺+1): 557.

EXAMPLE 120 Preparation of Compound 120:N-ethyl-N′-(4-{[2-(7-methyl-1H-indol-3-yl)-ethylamino]-methyl}-benzyl)-N-pyridin-2-ylmethyl-N′-{2-[(pyridin-2-ylmethyl)-amino]-ethyl}-ethane-1,2-diamine

Compound 120 was prepared in a manner similar to that described inExample 111 followed by reaction with acetaldehyde and NaBH₄ (Yield:75%).

LC-MS (M⁺+1): 590.

EXAMPLE 121 Preparation of Compound 121:3-{4-[(bis-{2-[(pyridin-2-ylmethyl)-amino]-ethyl}-amino)-methyl]-benzylamino}-1-(3-chloro-phenyl)-propan-1-ol

Compound 121 was prepared in a manner similar to that described inExample 16 (Yield: 70%).

LC-MS (M⁺+1): 573.

EXAMPLE 122 Preparation of Compound 122:1-(3-benzyloxy-phenyl)-3-{4-[(bis-{2-[(pyridin-2-ylmethyl)-amino]-ethyl}-amino)-methyl]-benzylamino}-propan-1-ol

Compound 122 was prepared in a manner similar to that described inExample 16 (Yield: 67%).

LC-MS (M⁺+1): 645.

EXAMPLE 123 Preparation of Compound 123:3-[2-((4-{[2-(7-methyl-1H-indol-3-yl)-ethylamino]-methyl}-benzyl)-{2-[(pyridin-2-ylmethyl)-amino]-ethyl}-amino)-ethylamino]-3-pyridin-2-yl-propan-1-ol

Compound 123 was prepared in a manner similar to that described inExample 111 (Yield: 73%).

LC-MS (M⁺+1): 606.

EXAMPLE 124 Preparation of Compound 124:3-[2-((4-{[(1H-benzoimidazol-2-ylmethyl)-amino]-methyl}-benzyl)-{2-[(pyridin-2-ylmethyl)-amino]-ethyl}-amino)-ethylamino]-3-pyridin-2-yl-propan-1-ol

Compound 124 was prepared in a manner similar to that described inExample 111 (Yield: 60%).

LC-MS (M⁺+1): 579.

EXAMPLE 125 Preparation of Compound 125:N-(4-{[(1H-benzoimidazol-2-ylmethyl)-amino]-methyl}-benzyl)-N′-pyridin-2-ylmethyl-N-{2[(pyridin-2-ylmethyl)-amino]-ethyl}-ethane-1,2-diamine

Compound 125 was prepared in a manner similar to that described inExample 16 (Yield: 70%).

LC-MS (M⁺+1): 535.

EXAMPLE 126 Preparation of Compound 126:N-(4-{[3-(3-chloro-phenyl)-propylamino]-methyl}-benzyl)-N′-pyridin-2-ylmethyl-N-{2-[(pyridin-2-ylmethyl)-amino]-ethyl}-ethane-1,2-diamine

Compound 126 was prepared in a manner similar to that described inExample 16 (Yield: 85%).

LC-MS (M⁺+1): 557.

EXAMPLE 127 in vitro Assay

Compounds 1-126 were tested for their efficacy in binding to CXCR4receptor using a DELFIA GTP-binding kit (Wallac Oy, Turku, Finland). TheDELFIA GTP-binding assay is a time-resolved fluorometric assay based onGDP-GTP exchange on G-protein subunits followed by activation of a Gprotein-coupled receptor by its agonists. Eu-GTP, obtained from WallacOy, was used in this assay to allow monitoring of agonist-dependentactivation of G-protein. Stimulation of CXCR4 receptor by SDF-1 leads tothe replacement of GDP by GTP on the α-subunit of G-protein. This GTP-Gαcomplex represents the activated form of G-protein. Eu-GTP, anon-hydrolysable analog of GTP, can be used to quantify the amount ofactivated G-protein. (Peltonen et al., Eur. J. Pharmacol. (1998)355:275.)

Plasma membrane of CXCR4-expressing HEK293 cells was suspended in anassay buffer (50 mM NaCl, 100 μg/mL saponin, 3 mM MgCl₂, 3 μM GDP, 5%BSA, 50 mM HEPES, pH 7.4). An aliquot (4 μg protein) was added to eachwell of an AcroPlate (Pall Life Sciences, Ann Arbor, Mich.). After theaddition of the test compounds (10 μM in 0.1% DMSO) and stromal-derivedfactor-1 (4 nM in the assay buffer), the assay plate was incubated inthe dark at room temperature with slow shaking for 10 minutes. Eu-GTPwas added to each well and the plate was incubated again for 60 minutes.The assay was terminated by washing the plate twice with a wash solutionprovided in the assay kit. Binding of Eu-GTP was determined based on thefluorescence signal from a Victor 2 multi-label reader.

Unexpectedly, all of the tested compounds showed IC₅₀ values lower than10 μM. Specifically, 97 of the test compounds showed IC₅₀ values lowerthan 1 μM. Among them, 56 showed IC₅₀ values between 0.004 μM and 0.1μM.

OTHER EMBODIMENTS

All of the features disclosed in this specification may be combined inany combination. Each feature disclosed in this specification may bereplaced by an alternative feature serving the same, equivalent, orsimilar purpose. Thus, unless expressly stated otherwise, each featuredisclosed is only an example of a generic series of equivalent orsimilar features.

From the above description, one skilled in the art can easily ascertainthe essential characteristics of the present invention, and withoutdeparting from the spirit and scope thereof, can make various changesand modifications of the invention to adapt it to various usages andconditions. Thus, other embodiments are also within the scope of thefollowing claims.

For example, the polyamine compounds described above can be used totreat an inflammatory or immune disease through mechanisms other thanbinding to CXCR4 receptor. Further, other uses of these compounds arealso within the scope of this invention.

1. A compound of the formula:

wherein X is —CH₂—, —C₂H₄—, or —C₃H₆—; Y is aryl, heteroaryl, C₃-C₈cycloalkyl, C₅-C₈ cycloalkenyl, C₃-C₈ heterocycloalkyl, or C₅-C₈heterocycloalkenyl; each of Z₁ and Z₂, independently, is —CH₂—, —C₂H₄—,or —C₃H₆—; R₁ is aryl or heteroaryl; R₂ is -A₁-B₁-D₁-E₁; R₃ is-A₂-B₂-D₂-E₂; and R₄ is -A₃-B₃-D₃-E₃ or, together with R₃, is C₄-C₂₀heterocycloalkyl; in which A₁ is —CH₂—, —C₂H₄—, —C₃H₆—, —C₄H₈—, or—C₅H₁₀—B₁ is —NR—; D₁ is —CH₂—, —C₂H₄—, or —C₃H₆—; and E₁ is aryl orheteroaryl; A₂ deleted; B₂ is deleted; D₂ is deleted; and E₂ is H, C₃-C₈cycloalkyl, C₅-C₈ cycloalkenyl, C₃-C₈ heterocycloalkyl, C₅-C₈heterocycloalkenyl, aryl, or heteroaryl; A₃ is —CH₂—, —C₂H₄—, —C₃H₆—,—C₄H₈—, —C₅H₁₀—, or deleted; B₃ is deleted; D₃ is deleted; and E₃ isC₃-C₈ cycloalkyl, C₅-C₈ cycloalkenyl, C₃-C₈ heterocycloalkyl, C₅-C₈heterocycloalkenyl, aryl, or heteroaryl; and each R, independently,being H or C₁-C₁₀ alkyl.
 2. The compound of claim 1, wherein X is —CH₂—or —CH(CH₃)—, Y is phenyl, Z₁ is —CH₂—, and Z₂ is —CH₂—.
 3. The compoundof claim 2, wherein R₃ is -A₂-B₂-D₂-E₂ or, together with R₄, is C₄-C₂₀heterocycloalkyl; A₁ is —C₂H₄— A₃ is —CH₂—, —C₂H₄—, —C₃H₆—, or deleted;B₁ is —NH—; D₁ is —CH₂— or —CH(CH₃)—; E₂ is H; and E₃ is aryl,heteroaryl, C₃-C₈ cycloalkyl, C₅-C₈ cycloalkenyl, or C₃-C₈heterocycloalkyl.
 4. A pharmaceutical composition comprising a compoundof the formula:

wherein X is —CH₂—, —C₂H₄—, or —C₃H₆—; Y is aryl, heteroaryl, C₃-C₈cycloalkyl, C₅-C₈ cycloalkenyl, C₃-C₈ heterocycloalkyl, or C₅-C₈heterocycloalkenyl; each of Z₁ and Z₂, independently, is —CH₂—, —C₂H₄—,or —C₃H₆—; R₁ is aryl or heteroaryl; R₂ is -A₁-B₁-D₁-E₁; R₃ is-A₂-B₂-D₂-E₂; and R₄ is -A₃-B₃-D₃-E₃ or, together with R₃, is C₄-C₂₀heterocycloalkyl; in which A₁ is —CH₂—, —C₂H₄—, —C₃H₆—, —C₄H₈—, or—C₅H₁₀—; is —NR—; D₁ is —CH₂—, —C₂H₄—, or —C₃H₆—; and E₁ is aryl, orheteroaryl; A₂ deleted; B₂ is deleted; D₂ is deleted; and E₂ is H, C₃-C₈cycloalkyl, C₅-C₈ cycloalkenyl, C₃-C₈ heterocycloalkyl, C₅-C₈heterocycloalkenyl, aryl, or heteroaryl; A₃ is —CH₂—, —C₂H₄—, —C₃H₆—,—C₄H₈—, —C₅H₁₀—, or deleted; B₃ is deleted; D₃ is deleted; and E₃ isC₃C₈ cycloalkyl, C₅-C₈ cycloalkenyl, C₃-C₈ heterocycloalkyl, C₅-C₈heterocycloalkenyl, aryl, or heteroaryl; and a pharmaceuticallyacceptable carrier.
 5. The composition of claim 4, wherein X is —CH₂— or—CH(CH₃)—, Y is phenyl, Z₁ is —CH₂—, and Z₂ is —CH₂—.
 6. The compositionof claim 5, wherein R₃ is -A₂-B₂-D₂-E₂ or, together with R₄, is C₄-C₂₀heterocycloalkyl; A₁ is —C₂H₄— A₃ is —CH₂—, —C₂H₄—, —C₃H₆—, or deleted;B₁ is —NH—; D₁ is —CH₂—or —CH(CH₃)—; E₂ is H; E₃ is aryl, heteroaryl,C₃-C₈ cycloalkyl, C₅-C₈ cycloalkenyl, or C₃-C₈ heterocycloalkyl.